JP2022511753A - Single dose use of a composition comprising a particular mixture of grape extract and blueberry extract - Google Patents

Abstract

The present invention relates to the use of a composition comprising at least one mixture of molecules obtained from Vitis vinifera and Vaccinium angustifolia, the mixture being at least 1% catechin and / or at least 1% catechin and / or given by weight relative to the total weight of the mixture. Epicatechin and-at least 5 ppm (1 million percent in the mixture) of ferulic acid and-at least 200 ppm of resveratrol are included as a single dose to improve or maintain cognitive function in humans or animals.

Description

The present invention relates to single dose use of a mixture of specific molecules for a rapid and immediate effect on cognitive function in humans and animals.

More and more healthy adults, especially college students, are looking for solutions to improve their cognitive abilities. This quest has led to an increase in the inadequate and sometimes dangerous use of pharmaceutical stimulants. In addition, existing commercial nutritional solutions for use in the context of chronic doses that last for several weeks can prove to be limited and expensive.

Therefore, for safe, non-pharmacological, natural, and non-restrictive solutions to improve cognitive performance, especially in the form of food supplements that are effective after a single dose, in public health. There is a need to arise from the benefits of. It is also important that the dose is in accordance with a small amount of the naturally active ingredient and is a single dose taken immediately prior to the desired effect, eg, prior to a laboratory test.

It is an object of the present invention to meet these needs. To this end, the present invention relates to the use of a composition comprising a mixture of molecules obtained from Vitis vinifera and Vaccinium angustifolia.

Mixtures of molecules particularly rich in flavonoids, obtained from Vitis vinifera and Vaccinium angustifolia, are particularly known from the filing of FR1560263, which have the effect of the mixture on human cognitive function during chronic ingestion. Explaining. Similarly, the contribution of flavonoid-rich grapes and blueberry / bilberry extracts has shown efficacy for memory when administered chronically for weeks in mice and dogs and months in humans.

However, flavonoids are of interest to cognition when taken chronically, but when they are taken in single doses, they do not always have an effect and their activity is considered flavonoids. Very variable, depending on the type and dose (often very large), and the age of the people involved. Therefore, two clinical studies conducted with cocoa beverages in acute doses (single doses) showed beneficial effects of high doses of cocoa flavanols (520, 773, and 994 mg), but 46 mg of flavanols. The controls provided had no effect (Field DT, Williams CM, Butler LT. Consumption of cocoa flavanols results in an acte improvement invisual aviment invisual and gen. 60, Scholey AB, French SJ, Morris PJ, Kennedy DO, Milne AL, Haskell CF.Consumption of cocoa flavanols results in acute improvements in mood and cognitive performance during sustained mental effort.J Psychopharmacol.2010 Oct; 24 (10): 1505 -14).

Pase et al. (Pase MP, School AB, Pipenas A, Kras M, Nolidin K, Gibbs A, Wesnes K, Studio C. Cocoa polyphenols enhance recognition associated with acute consumption of cocoa flavonoids) -Another study by controlled trial. J Polyphenol. 2013 May; 27 (5): 451-8) also shows no improvement in cognitive function. Similarly, Wightman et al., 2012 (Wightman EL, Haskell CF, Forster JS, Veasey RC, Kennedy DO.Epigallocatechin gallate, cerebral blood flow parameters, cognitive performance and mood in healthy humans: a double-blind, placebo-controlled, crossover investigation Hum Psychopharmacol. 2012 Mar; 27 (2): 177-86) examined the cognitive effects of green tea extract at doses of 135 mg and 270 mg of flavonoids on cognition, with no significant results reported. did. Finally, two randomized, controlled (placebo), double-blind studies, Hendrickson and Mattes (Hendrickson SJ, Mattes RD. No acte effects of grape juice on application, memory. Memory. 52), and Haskell Ramsay et al (Haskell-Ramsay CF, Stuart RC, Okello EJ, Watson AW.Cognitive and mood improvements following acute supplementation with purple grape juice in healthy young adults.Eur J Nutr.2017 Dec; 56 (8): 2621-2631) studied the acute effects of grape juice consumption on cognition (episode memory, work memory, attention) in young adults aged 21 and 26, respectively, on average. In the first study, 230 ml of grape juice providing 138 mg of anthocyanin and 1504 mg of polyphenol was consumed, and in the second study, 600 ml of grape juice containing 580 mg of anthocyanin and 2100 mg of polyphenol was supplied to each volunteer. Both of these studies showed no effect on memory.

It is also very surprising that a composition that has a cognitive effect on chronic ingestion also has a cognitive effect on a single dose. FIG. Targeting the use of the mixture, the mixture is
-With at least 1% catechin and / or epicatechin, the percentage given by weight to the total weight of the mixture.
-At least 5 ppm (parts per million in the mixture) of ferulic acid and
-Contains at least 200 ppm resveratrol.

Advantageously, a single dose of such a composition makes it possible to obtain maintenance or improvement in cognitive performance in less than 5 hours with low doses and natural products.

With reference to the accompanying drawings, the following detailed description of the invention reveals other features and advantages.

-FIG. 1 shows a plan of clinical trials conducted to assess efficacy according to the invention. -FIG. 2 is a graph showing changes in capacity compared to B1 in repetitive batteries (B2-B6), grouped by treatment (P: placebo, V: research drug (Verum)). The change in the "net" score (correct answer-wrong answer) in the minus 3 subtraction test is shown in A. This same test shows% variation in correct response (B) and number of responses per block (C). The result of the correct answer for the rapid visual information processing test is shown in D. The p-value is the value of the corresponding t test. The vertical bar represents the standard deviation. -Figure 3 shows the synthetic Z-score of the correct answers in the STS + SSS + RVIP test, i.e., this data represents the percentage variation in the number of cumulative correct answers compared to B1 grouped by treatment (placebo, invention) (placebo, invention). A). Battery analysis was performed using the corresponding t-test. The vertical bar represents the standard deviation. The development within the treatment (placebo, invention) is evaluated by multiple comparisons and the results are found in Table B. -Fig. 4 shows the effect of treatment on the dilated peak of the brachial artery (60 seconds after ischemia) mediated by blood flow. There is no difference in the placebo vs. invention correspondence t-test.

Definitions In the sense of the invention, "animal" means any animal that can receive the composition according to the invention, such as, but not limited to, pets, poultry, pigs, ruminants, goats, or even. Means mouse.

In the sense of the present invention, "anthocyanidin" means all anthocyans or anthocyanins or anthocyanids in an aggrecorn or glycosylated form (ie, carrying a sugar). Therefore, in the present application and within the meaning of the present invention, the terms "anthocyanidin", "anthocyan", "anthocyanin", and "anthocyanid" are equivalent.

In the sense of the present invention, "at least X% of catechins and / or epicatechins" means at least X% of catechins in the absence of epicatechins in the mixture, or at least X in the absence of catechins in the mixture. % Epicatechin, or a mixture of at least X% catechin and epicatechin, if both catechin and epicatechin are present in the mixture. Preferably, the aforementioned word means a mixture of at least X% catechin and epicatechin.

In the sense of the present invention, "an extract of Vaccinium angustifolia" means at least one molecule, preferably a set of molecules, obtained from Vaccinium angustifolia. The raw material may be leaves and / or fruits, preferably all raw materials are plant leaves and fruits.

In the sense of the present invention, "Vitis vinifera extract" means at least one molecule, preferably a set of molecules, obtained from Vitis vinifera. The raw material may be leaves and / or fruits and / or seeds and / or pericarp, xylem, preferably the raw materials are the above-ground parts of the plant, i.e. leaves, fruits, bark (ie, pericarp), seeds and. All of the xylem, and even more preferably the pericarp (skin) and seeds. The combination of resveratrol-rich pericarp with flavanol monomers, procyanidin oligomers, and proanthocyanidin-rich seeds may be particularly advantageous for the present invention.

In the sense of the present invention, "an extract obtained from a mixture of Vicis vinifera and Vaccinium angustifolia" means a set of molecules obtained from a mixture of Vitis vinifera and Vaccinium angustifolia. The raw material for Vitis vinifera may be leaves and / or fruits and / or seeds and / or xylem, preferably the raw material for Vitis vinifera is the above-ground part of the plant, ie leaves, fruits, skins (pericarp),. All of the seeds and xylem, more preferably the pericarp (skin) and seeds. The raw material for Vaccinium angustifolia may be leaves and / or fruits, preferably the raw material for Vaccinium angustifolia is all plant leaves and fruits.

Within the meaning of the present invention, the term "flavanol polymer" means flavanols exhibiting a degree of polymerization greater than 10.

Within the meaning of the present invention, the term "ppm" means parts per million (mg / kg) in a mixture. Unless otherwise indicated, ppm refers to the weight relative to the total weight of the mixture.

In the sense of the present invention, "single dose" means the consumption of one or more consumption units taken at one time.

In the sense of the present invention, "healthy" is non-pathological, unaffected by pathology, especially neurological pathology, but may be tired and / or stressed. Means a human or animal in a state.

Accordingly, the present invention relates to the use of a composition comprising at least one mixture of molecules obtained from Vitis vinifera and Vaccinium angustifolia as a single dose in healthy humans or animals to improve or maintain cognitive function. teeth,
-A percentage of at least 1%, preferably at least 5%, even more preferably 5% to 50%, particularly 7% to 35%, is given by weight to the total weight of the mixture, catechins and /. Or with epicatechin,
-With ferulic acid at least 5 ppm (1 million percent in the mixture), preferably at least 10 ppm, even more preferably 5 ppm to 300 ppm, particularly 10 ppm to 100 ppm.
-Contains at least 200 ppm, preferably at least 300 ppm, even more preferably at least 400 ppm, even more preferably 300 ppm to 6000 ppm, particularly 400 to 6000 ppm resveratrol.

The composition is also used as a single dose for an acute effect for acute treatment, especially in sick humans or sick animals suffering from diseases that can affect cognitive performance, such as colds. You can also do it. Accordingly, the present invention is to be used as a single dose, essentially a single non-repeating base, to improve or maintain cognitive function in a diseased human or animal. With respect to a composition comprising at least one mixture of molecules obtained from, the mixture is also:
-A percentage of at least 1%, preferably at least 5%, even more preferably 5% to 50%, particularly 7% to 35%, is given by weight to the total weight of the mixture, catechins and /. Or with epicatechin,
-With ferulic acid at least 5 ppm (1 million percent in the mixture), preferably at least 10 ppm, even more preferably 5 ppm to 300 ppm, particularly 10 ppm to 100 ppm.
-Contains at least 200 ppm, preferably at least 300 ppm, even more preferably at least 400 ppm, even more preferably 300 ppm to 6000 ppm, particularly 400 to 6000 ppm resveratrol.

This use is a single dose and is carried out 5 hours before the desired effect.

Mixtures of molecules of useful compositions according to the invention are also preferably at least 1% catechin and / or epicatechin plus at least 5 ppm ferulic acid and at least 200 ppm resveratrol:
-At least 50 ppm quercetin and / or quercetin glycosides, preferably at least 70 ppm, particularly 50 ppm to 10,000 ppm quercetin and / or glycosides, and / or-at least 500 ppm, preferably at least 600 ppm, even more. It preferably contains at least 700 ppm, and even more preferably 600 ppm to 5000 ppm of anthocyanidins.

Preferably, the malvidin 3 glucoside is the predominant anthocyanidin. Preferably, it is present in the mixture at a concentration of at least 300 ppm.

According to the first embodiment, the mixture of molecules is a mixture consisting of an extract of Vitis vinifera and an extract of Vaccinium angustifolia.

According to the second embodiment, the mixture of molecules is a mixture consisting of an extract obtained from a mixture of Vitis vinifera and Vaccinium angustifolia.

According to the third embodiment, the mixture of molecules is
-Extracts of Vitis vinifera and extracts of Vaccinium angustifolia,
-A mixture consisting of an extract obtained from a mixture of Vitis vinifera and Vaccinium angustifolia.

Preferably, the Vitis vinifera extract present in the useful composition according to the invention has a flavanol polymer content of less than 0.5% by weight of the total weight of the polyphenols in the extract, even more preferably 0.1. An extract with a content of less than%. According to the present invention, flavanol polymers are less bioavailable, unlike flavanol monomers, which are very rapidly absorbed into the small intestine and then metabolized to methylated, sulfated, and gluclonized derivatives. This low presence of polymer is a quality standard for grape extracts used specifically for efficacy and bioavailability.

The extract according to the present invention is
-Catechin and catechins in an amount of at least 1% by weight, preferably at least 5% by weight, even more preferably 5% to 50% by weight, particularly 7% to 35% by weight, based on the total weight of the mixture. / Or with epicatechin,
-With ferulic acid at least 5 ppm, preferably at least 10 ppm, even more preferably 5 ppm to 300 ppm, especially 10 ppm to 100 ppm.
-With resveratrol at least 200 ppm, preferably at least 300 ppm, even more preferably at least 400 ppm, even more preferably 300 ppm to 6000 ppm, especially 400 to 6000 ppm.
-Optionally, with at least 50 ppm quercetin and / or quercetin glycosides, preferably at least 70 ppm, particularly 50 ppm to 10,000 ppm quercetin and / or glycosides.
-Any option that makes it possible to obtain a mixture comprising at least 500 ppm, preferably at least 600 ppm, even more preferably at least 700 ppm, even more preferably 600 ppm to 5000 ppm of anthocyanidins. It can be obtained by the method.

Preferably, malvidin 3 glucoside is the major anthocyanidin at a concentration of at least 300 ppm. Preferably, the anthocyanidin comprises at least 20%, more preferably at least 25% malvidin 3 glucoside (percentage by weight).

Particularly preferred methods include the following steps:
-Obtaining an extract of Vitis vinifera:

* Perform water and / or ethanol extraction of all leaves, fruits, pericarps, seeds, and xylem of Vitis vinifera, preferably the amount of solvent used (30% v / v-96%). V / V) is 2 to 10 times the mass of the material used. The extraction time can be 30 minutes to 24 hours and the extraction temperature can be 20 ° C to 80 ° C. The raw materials used can be dried, fresh, or frozen, as is, or crushed.
* Separation of water and / or ethanol solution from solid materials, for example by centrifugal decanting, or by squeezing and filtration;
* Evaporating ethanol, preferably by vacuum evaporation at a temperature below 60 ° C. and a pressure below 100 mbar;
* Preferentially select proanthocyanidin monomers and oligomers (including those with a degree of polymerization of 2-10) to eliminate flavanol polymers (> decamers) and to the total weight of polyphenols in the extract. In contrast, membrane separation of the pre-dissolved extract is performed in order to obtain an extract characterized by a flavanol polymer content of less than 0.5% by weight, more preferably less than 0.1% by weight. , 15,000 daltons, more preferably less than 3000 daltons, can be performed using a filtration membrane with a cutoff threshold.
-Obtaining an extract of Vaccinium angustifolia:
* Perform water and / or ethanol extraction of all leaves and fruits of Vaccinium angustifolia, preferably Vaccinium angustifolia; the amount of solvent used (30% v / v-96% V / V) is used. It is 2 to 10 times the mass of the material. The extraction time can be 30 minutes to 24 hours and the extraction temperature can be 20 ° C to 80 ° C. The raw materials used can be dry, fresh, or frozen;
* Separation of water and / or ethanol solution from solid material by centrifugal decanting or by squeezing and filtration;
* Evaporating ethanol, preferably by vacuum evaporation at a temperature below 60 ° C. and a pressure below 100 mbar;
* Drying the extract by micronization, vacuum oven, or lyophilization with or without a support such as maltodextrin;
-Mixing the extract of Vitis vinifera with the Vaccinium angustifolia before or after the drying step.

According to one variant, the method involves performing the following steps:
-Mixing Vitis vinifera and Vaccinium angustifolia to carry out water and / or ethanol extraction of all leaves and fruits of Vaccinium angustifolia, preferably the amount of solvent used (30% v / v). 96% V / V) is 2 to 10 times the mass of the material used. The extraction time can be 30 minutes to 24 hours and the extraction temperature can be 20 ° C to 80 ° C. The raw materials used can be dry, fresh, or frozen;
-Separate water and / or ethanol solutions from solid pomace by centrifugal decanting or by squeezing and filtration;
-Evaporating ethanol by vacuum evaporation, preferably at a temperature below 60 ° C. and a pressure below 100 mbar;
-Drying the extract by spraying or sublimation with or without a support such as maltodextrin.

Regardless of the modification of the method, before the drying process, the method is as follows:
-Filling the resin with a solution of mixed or unmixed extract,
-Rinse the resin with water,
-Applying a water / ethanol elution solution to the resin,
-Recovering the purified eluate,
-Evaporating ethanol from the eluate,
-Concentrating the eluate,
-The purified aqueous extract can be dried.

Compositions according to the invention can exclusively consist of a mixture of molecules, i.e., an extract or contain other components. Preferably, in addition to the mixture of molecules, the composition according to the invention contains other components, in particular excipients or coatings such as maltodextrin, microcrystalline cellulose, cyclodextrin, starch, soluble or insoluble fibers. ..

The composition may be in any form suitable for nutritional application, preferably in powder form.

The composition is in the form of another composition selected from tablets, capsules, gelatin capsules, powders, solutions, microcapsules, suspensions, emulsions, food supplements, beverages and foods, in particular for humans or animals. It can be incorporated into the provided nutritional composition.

The composition is a non-therapeutic nutritional composition intended for humans, such as food supplements, bars, dairy products, swallowed or rehydrated powders, gels, jams, candies, carbonated or non-carbonated beverages. , A dry beverage to be rehydrated, preferably a component.

The composition can also be a nutritional composition intended for animals.

The composition is also a dry food such as a kibble (extruded, co-extruded, or freeze-dried), a wet or semi-wet food such as a bite in a sweet, a snack, a sauce, a bite in a jelly, a beverage, a beverage. , Or even a non-therapeutic nutritional composition for animals, such as food supplements. Preferably, the composition according to the invention is incorporated into a dry food such as kibble.

Advantageously, when the composition according to the invention is directed to an animal, the composition is in the composition, especially in the nutritional composition, as an inclusion, eg, by impregnation or mixing, or by coating. Incorporated by addition to the entity, i.e., by premixing with one or more components, such as, for example, at least one palatability enhancer, by applying it to the surface of the composition by spraying or dusting. be able to.

It can be a pharmaceutical composition, drug, or veterinary product.

The presence of molecules of Vitis vinifera and Vaccinium angustifolia specific to the mixture present in the compositions according to the invention allows for synergistic effects in the maintenance or amelioration of cognitive function.

Therefore, a particular object of the invention is the composition described above for use as a single dose in healthy humans or animals to improve or maintain cognitive function, regardless of its variant.

The single dose is taken less than 5 hours before the desired effect, preferably less than 3 hours before the desired effect. This single dose can be taken once on a non-repeating basis.

Preferably, the invention is to improve or maintain cognitive function, i.e., in particular, memory and / or executive function and / or attention and / or concentration and / or learning and / or flexibility and / or planning and / Or with respect to use as a single dose to improve or maintain agility.

In particular, the compositions according to the invention can be used in humans to improve or maintain performance in tests or tests that require sustained cognitive ability.

The compositions according to the invention, taken as a single dose in humans, maintain or improve cognitive function in humans, preferably over 10 years of age, even more preferably over 15 years of age, even more preferably over 18 years of age. It is especially useful and effective for doing so. These include college students and active adults.

The compositions according to the invention, taken as a single dose in an animal, are particularly useful and effective for maintaining or ameliorating cognitive function in dogs, cats, or horses.

Preferably, the composition according to the invention is for humans or animals.
-At least 100 μg / kg body weight, preferably 500 μg / kg, even more preferably 1 mg / kg, catechin and / or epicatechin, and-preferably at least 0.05 μg / kg body weight, preferably. 0.25 μg / kg, even more preferably 0.5 μg / kg of ferulic acid, and − at least 10 μg per kg of body weight, preferably 50 μg / kg, even more preferably 100 μg / kg, resveratrol. And-optionally at least 0.2 μg / kg body weight, preferably 1 μg / kg, even more preferably 2 μg / kg, quercetin and / or quercetin glycoside, and-optionally, per kg body weight. It is used in an amount that makes it possible to provide at least 1 μg, preferably 5 μg / kg, and even more preferably 10 μg / kg of anthocyanidin.

Here, the invention is described through examples and test results demonstrating a synergistic effect at a single and rapid dose on cognitive function of the compositions of interest in the present application.

Example 1: Composition according to the present invention This first example of the mixture according to the present invention is obtained by carrying out the method described below.

The raw materials used are:
-Fruit skins and seeds of Vitis vinifera, a by-product of the wine industry,
-Frozen berries of Vaccinium angustifolia.

400 g of frozen berries of Vaccinium angustifolia are ground and mixed with a solution of 2000 ml of 80% (V / V) ethanol with a content of 0.1 wt% HCl. The mixture is kept at room temperature (20 ° C.) for 24 hours. The ethanol solution is then separated from the pulp by filtration and concentrated under vacuum on a rotary evaporator with 20% dry material. Some of this extract is stored for testing and the other is stored for mixing with the extract of Vitis vinifera.

500 g of Vitis vinifera pericarp and Vinifera seeds are mixed with 2500 ml of 80% (V / V) ethanol at 40 ° C. for 5 hours with a content of 0.1 wt% HCl. The ethanol solution is then separated from the pulp by filtration. Ethanol is then removed under vacuum on a rotary evaporator at a temperature of 50 ° C. under 60 mbar. The aqueous solution is then diluted to give a 5% dry material and filtered through a 5000 Dalton membrane. Then, the obtained penetrant is filled in the resin column (C18) at 1 BV / hour. The resin is then first rinsed with 3BV distilled water at 2BV / hour and then eluted with a 5BV 80% (V / V) ethanol solution at 1BV / hour.

A portion of the extracted solution is tested and characterized (Table 1a).

The polyphenols presented in this table were measured by ultra-high performance liquid chromatography using a fluorescence detector.

The other moieties are then mixed with an extract of Vaccinium angustifolia to form a mixture according to the invention and maltodextrin is added to the mixture until a solution with a solid content of 30% is obtained.

The solution is then spray dried at an inlet temperature of 160 ° C.

The product obtained is a mauve powder containing the polyphenols shown in Table 1b. The polyphenols presented in this table were measured by mass / mass ultrafast liquid chromatography (UPLC-MS / MS) using a fluorescence detector.

Example 2: Composition According to the Invention This second example of the mixture according to the invention is obtained by carrying out the method described below.

The raw materials used are:
-Vitis vinifera seeds and pericarp,
-Vaccinium angustifolia berries.

1000 g of frozen Vaccinium angustifolia pomace is ground and mixed with 5000 ml of a solution of 60% (V / V) ethanol with a content of 0.1 wt% HCl. The mixture is kept at room temperature (20 ° C.) for 24 hours. The ethanol solution is then separated from the pulp by filtration and concentrated under vacuum on a rotary evaporator with 20% dry material.

400 g of selected Vitis vinifera peels and seeds are mixed at 60 ° C. with 1500 ml of 80% (V / V) ethanol for 5 hours. The ethanol solution is then separated from the pulp by filtration. Ethanol is then removed under vacuum on a rotary evaporator at a temperature of 50 ° C. under 60 mbar. The aqueous solution is then diluted to give a 5% dry material and filtered through a 5000 Dalton membrane. Then, the obtained penetrant is filled in the resin column (C18) at 1 BV / hour. The resin is then first rinsed with 3BV distilled water at 2BV / hour and then eluted with a 50% (V / V) ethanol solution of 5BV at 1BV / hour.

A portion of the extracted solution is tested and characterized (Table 2a).

The product obtained is a mauve powder containing the polyphenols shown in Table 2b. The polyphenols presented in this table were measured by ultra-high performance liquid chromatography (UPLC-MS / MS) using a fluorescence detector.

Example 3: Compositions According to the Invention with Excipients for Humans 19.980 kg of the composition of Example 1 is mixed with 0.020 kg of colloidal silica. The composition is obtained by mixing the constituents under conventional conditions known to those of skill in the art. The composition is packed in a PET bag, which itself is packed in a cardboard box.

Example 4: Nutritional composition according to the present invention for humans The raw materials used are as follows:
-Fresh grapes of Vitis vinifera used for wine making,
-Frozen berries of Vaccinium angustifolia.

7.2 g of Vaccinium angustifolia frozen berries are ground and squeezed. The juice is separated from the solid fraction (1.8 g). The solid fraction is mixed with a solution of 35 ml of 80% (V / V) ethanol with a content of 0.1 wt% HCl. The mixture is kept at room temperature (20 ° C.) for 24 hours. The ethanol solution is then separated from the pulp by filtration and concentrated under vacuum on a rotary evaporator with 20% dry material. The extract is then dried under vacuum on a maltodextrin support. 180 mg of the extract in powder form is then harvested and mixed with the extract of Vitis vinifera.

300 g of fresh grapes are squeezed and the juice is separated from a solid substance (71 g) consisting of the peel and seeds of Vitis vinifera. The solid material is mixed with 1000 ml of 80% ethanol (V / V) at 40 ° C. for 5 hours with a content of 0.1 wt% HCl. The ethanol solution is then separated from the pulp by filtration. Ethanol is then removed under vacuum on a rotary evaporator at a temperature of 50 ° C. under 60 mbar. The aqueous solution is then diluted to give a 5% dry material and filtered through a 5000 Dalton membrane. Then, the obtained penetrant is filled in the resin column (C18) at 1 BV / hour. The resin is then first rinsed with 3BV distilled water at 2BV / hour and then eluted with a 5BV 80% (V / V) ethanol solution at 1BV / hour. The aqueous alcohol solution is then evaporated on a rotary evaporator at a temperature of 50 ° C. at 60 mbar and then the resulting extract is dried under vacuum. Then 420 mg of dried Vicis vinifera extract is recovered and then mixed with 180 mg of Vaccinium angustifolia extract to make up 600 mg of the product according to the invention.

The product according to the invention is administered in the form of gelatin capsules. Each gelatin capsule contains a total of 450 mg of powder consisting of 300 mg of a product according to the invention supplemented with 150 mg of corn maltodextrin.

The recommended daily product dose is 600 mg, or 2 gelatin capsules.

Example 5: Nutritional composition according to the present invention for humans Example 5
Composition of Example 1: 300 m
-Vitamin C: 80 mg
-A 400 mg gelatin capsule consisting of maltodextrin: 20 mg.

The composition is obtained by mixing the constituents under conventional conditions known to those of skill in the art and is also placed in gelatin capsules under conventional conditions.

The recommended amount is 1-2 gelatin capsules per day.

Example 6: Nutritional Composition of the Present Invention for Animals The composition of the present invention of Example 2 is compliant with AFCO standards and contains animal diets, fats, fibers, cereals and preservatives and antioxidants. Added to extruded dry dog food containing.

Addition of the composition to the kibble was carried out according to some embodiments, especially by coating and inclusions.

A coating test was performed by adding a composition according to the present invention to a liquid D'Tech poultry palatability enhancer (SPF, Elven, France). A first layer of poultry fat (6% by weight of the kibble) was added as a coating on the kibble, followed by a palatability enhancer (1%, 2% or 3%,% by weight of the kibble). A layer of mixture between the composition according to the invention (0.02%, 0.04% or 0.1%,% is relative to the weight of the kibble) was added.

Prior to extrusion, a composition according to the invention (0.02%, 0.04% or 0.1%,% is relative to the weight of the kibble) is added to the raw material (also referred to as premix) by adding. An inclusion test was performed.

Evaluation of the effect of a single dose on cognitive and endothelial function of the composition according to the invention A single-center, crossover, double-blind, placebo-controlled clinical trial was conducted in 30 healthy student volunteers aged 18-25 years. It was carried out against.

The composition of the product under study is the composition of Example 4.

The purpose of this study is, on the one hand, for cognitive function (working memory and attention) measured by tests conducted using a validated computer battery (COMPASS), and on the other hand, blood flow-dependent dilation (). Focused on the effect of a single dose of the product according to the invention on the endothelial function assessed by measuring FMD).

In fact, it has been found that the major known mechanism of action of the composition's effectiveness on cognitive function is associated with improved endothelial function, more specifically increased availability of nitric oxide (NO). Has been done. By either increasing synthesis by endothelial cells or limiting their degradation (enzymes, reactive oxidants). NO increases cerebral blood flow and therefore plays a major role in local vasodilation during neuronal activation. A simple non-invasive method for assessing endothelial function consists of measuring blood flow-dependent brachial artery diameter dilation (FMD) in response to transient ischemia.

The composition of Example 4 was administered in a single oral dose of two gelatin capsules (300 mg / capsule, ie 600 mg / dose). Placebo was a capsule of similar appearance containing only maltodextrin (without polyphenols). The research plan is shown in Fig. 1.

The subject had to comply with the following instructions.
-During the test: Do not change daily habits, especially those related to food, sleep, physical and intellectual activity. Do not take food supplements. Do not quit smoking or take drugs. Fill in the daily sleep time log.
-From 24 hours prior to each assessment visit (V1 and V2): Comply with dietary restrictions that allow mitigation of polyphenol consumption from 24 hours prior to the study and complete a dietary follow-up log.
-Do not engage in intense physical activity for more than 2 hours the day before the evaluation visit.
-Evaluation 12 hours fast before visiting the hospital.
-Bring dietary surveys and sleep tracking logs to the evaluation visit.

After confirming inclusion and exclusion criteria for cognitive assessment, subjects entered the study and received a training session on a computer cognitive assessment battery set up for this study using a module of COMPASS software (Northumbria University, Newcastle). The purpose of this battery was to model the conditions for school / university exams. To do this, we used a series of long, cognitively demanding, computerized tests. It consists of 6 series of batteries, each consisting of: 2 minutes continuous 3 subtraction (STS), 2 minutes continuous 7 subtraction (SSS), 5 minutes quick visual information Processing (RVIP), and subjective assessment on a 2-minute visual analog scale (performance, anxiety, performance, fatigue). Details of goals and workflows are listed below:

Continuous 3 subtraction (STS):
This task tested the subject's attention and working memory. This consists of asking participants to perform the subtraction of -3 and fill in the answer using the computer keyboard as quickly and accurately as possible. First, a randomly drawn number from 800 to 999 is displayed on the screen. This number disappears when the participant enters the first entry. If the answer is incorrect, the next correct answer will be automatically determined based on the last answer entered by the participants. The task lasts 2 minutes.

Continuous 7 subtraction (SSS):
This task also evaluated the subject's attention and working memory. The operation of this task is the same as STS, with the difference that participants are required to perform a -7 subtraction. The task lasts another 2 minutes.

Rapid Visual Information Processing (RVIP) Tasks:
This task aims to measure a participant's attention period. Throughout the test, numbers were displayed one by one on the screen (100 / min). Participants were asked to detect the appearance of three consecutive even or odd numbers and to press the spacebar on the keyboard as quickly as possible if a sequence of numbers was detected. The task lasts 5 minutes.

The total duration of this battery with high cognitive demand was 66 minutes. During these 66 minutes, the subject performs a series of 6 iterations of the battery, including STS, SSS, RVIP, and then subjective evaluation. Each battery lasts 11 minutes.

Endothelial function evaluation FMD measurement procedure Target preparation:
Research participants declared that they had fasted for 12 hours and followed the instructions. For each participant, FMD measurements were taken simultaneously during both visits.

Facility:
Recordings were performed by the same operator in a dedicated room using an ultrasonic machine (VINNO ™ E10) and a linear probe (VINNO ™ F4-42L) at 10 MHz.

Image acquisition:
Subjects were guided to the FMD laboratory by a clinical research technician, then placed in a health examination bed and rested comfortably on their backs for at least 15 minutes. After measuring the blood pressure, the right arm was fixed. A blood pressure cuff was attached to the forearm above the wrist. The recording probe was placed just above the wrinkles on the elbow of the subject's arm and secured by a retention system to obtain a clear image of the brachial artery (maximum diameter) without reorienting the probe.

Blood flow-dependent dilation: FMD
Longitudinal images of the arteries were recorded for 2 minutes (divided into 4 consecutive 30 seconds results due to the limitations of the possible recording time by the ultrasound machine). The operator then inflated the blood pressure cuff to 230 mmHg above systolic blood pressure for 6 minutes to induce a transient ischemic phase. Immediately after releasing the cuff compression, arterial images were re-recorded for 2 minutes (divided into 4 consecutive 30-second sensations).

Same subject, same during subsequent measurements by measuring the distance between the position of the probe and the wrinkles of the elbow in centimeters at the end of the ^{first ultrasonography at the first visit} (V1) Brachial artery recording could be performed at the position, improving reproducibility.

Record Analysis Procedure Records were analyzed using the Vascular Research Tools software with a semi-automatic vessel wall detection system and the Brachyal Analyzer module. All analyzes were performed by the same operator who was blind to the harvested product.

FMD was calculated as a percentage increase in arterial diameter from its baseline value according to the following equation: FMD = (maximum diameter-basal diameter) / basal diameter x 100. Basilar artery diameter values were calculated as the average of the values obtained during the 2 minutes prior to compression. The maximum diameter value was calculated as the highest average of the four values (30 seconds on average) measured after the release of compression.

Statistical analysis Analysis of cognitive test data Scores and homogeneity of arbitrary treatment / sequence interactions were determined by performing repeated measures ANOVA (rANOVA) for _each test at B1.

For each type of test, changes in performance over 6 iterations of the battery were estimated by performing rANOVA on score variability (total variation in the number of correct responses compared to B1).

Various tests to assess attention and working memory using composite scores, or "Z ⁱ composite" scores: a single measurement was obtained taking into account all correct scores obtained with STS, SSS, and RVIP. .. The ^Zi complex has made it possible to quantify the overall cognitive performance of each individual and each battery (block). In order to obtain the Z ⁱ complex of subject "i", the results of each test were centered and reduced to obtain the distance from the group mean as a function of standard deviation (Z score). This showed Z ⁱ _STS , Z ⁱ _SSS , and Z ⁱ _RVIP . This normalization step makes it possible to calculate the average of the tests that are grouped together, referred to herein as Z ⁱ complex [(Z ⁱ _STS + Z ⁱ _SSS + Z ⁱ _RVIP ) / 3)].

FMD Data Analysis Ultrasound reproducibility was evaluated by calculating intra- and inter-visit variability. Quantitative variables were explained by calculating the mean and standard deviation. The effectiveness of treatment was assessed by performing Student's t-test (paired and unpaired, depending on the condition).

All statistical tests related to cognitive tests were performed using R (Jupyter.org support). Tests on FMD data were performed using Prism6 software (GraphPad). All statistical studies performed in this study were bilateral with a confidence interval of 5%.

Results Group characteristics The initial characteristics of the 26 subjects in the ITT population are shown in Table 5 below.

Within this ITT population, the values of the FMD analysis of the four subjects were not interpretable in all respects. Therefore, these four subjects were excluded from the ITT population for FMD statistics, and therefore the number of subjects with similar characteristics to the total ITT population was set to 26 (Table 6).

Evolution of scores over time (STS, SSS, RVIP) Evolution of cognitive performance over battery testing was monitored via variation in net score (correct-wrong) _per block compared to B1 (FIG. 2A). Over the course of repeated STS tasks, the subject's STS score increased with the present invention, significantly increasing from _B4 , but not with placebo. A more detailed analysis of the results showed that this improvement in net performance was in fact associated with maintaining the number of correct answers throughout the test, coupled with the increased number of tasks under the composition according to the invention (Fig. 2C). It was shown that (Fig. 2B). Despite the visual appearance that may be preferred for the treatment of the present invention, no significant results were shown with SSS and RVIP (Fig. 2D).

Evolution of "overall performance" over time The Zi composite method was used to obtain an overall score of correct scores in the attention and working memory tests performed (STS, SSS, ^RVIP ). After normalization, the combination of different results showed a significant difference in overall performance (Fig. 3A). _In fact, for B4 to _B6 , the score with the study drug was significantly higher than that with placebo. In addition, by comparing evolution within treatment ( _bonferroni block vs. block), the score under placebo was significantly reduced, while under study drug, overall performance levels remained at _B1-6 . Note that this was done (Fig. 3B).

Results of Endothelial Function Quality of FMD Recording Variability within individual visits: Calculated by taking the first basal diameter for the same subject compared to the second basal diameter of the visit. The individual values are then averaged to obtain the individual mean intra-visit variability observed at 1.21 ± 5.2% (52 values).

Variability between individual visits: Calculated by taking the first basal diameter of V1 for the same subject compared to the same measurement of V2. The individual values are then averaged to obtain the mean inter-visit variation observed at 0.51 ± 4.14% (26 values).

Acute effects of the present invention on FMD An average of 5.93% under the present invention and an average of 5.73% (p = 0.45) under placebo, during treatment on the FMD peak (60 seconds after ischemia). No statistically significant difference was observed. The overall magnitude of dilation after ischemia did not differ significantly with treatment (Fig. 4).

Closing the study This study was conducted in healthy students (young adults), and single-dose supplementation with the compositions according to the invention maintains cognitive performance during long-term, cognitively demanding trials. It was possible to evaluate the effect on. Surprisingly, the combination and specific amounts of polyphenols present in the compositions according to the invention make it possible to obtain acute effects on cognition.

The results of this study show that the results of working memory tests are improved by the present invention through "qualitative" (more accurate answers) and "quantitative" (more attempts) improvements. In comparison, the same subject on placebo showed a decrease in working memory and attention performance over repeated test batteries. Therefore, single dose administration of the composition according to the invention allows for improved overall performance during long-term cognitive challenges (test simulations). Interestingly, about 2 hours after taking the product, the effect on cognitive function was maximized with battery B4.

However, no simultaneous significant changes in endothelial function were observed under these same conditions. Therefore, this effect is not associated with improved perfusion of brain structures involved in performing the tasks of these studies.

Therefore, the results of this study on the effect of a single dose on cognitive function and the rapid effect of the composition appear to be unpredictable, and those skilled in the art will also observe the effect of the product on FMD compared to placebo. You would have expected it to be.

actual:
-The test dose of flavonoids (≧ 174 mg) was lower than the minimum dose (520 mg cocoaoflabonoids) for which acute effects on cognitive function were demonstrated in the prior art.
-The compositions according to the invention did not induce a significant effect on FMD as compared to placebo.
-Previous studies on this product do not support the hypothesis of significant antioxidant activity after a single dose.
-Other potential mechanisms of action of this product (actions on neurogenesis and neuroplasticity) involve long processes and are unlikely to have significant physiological effects after just a few hours.

Several clinical studies conducted on products made from blueberries or grapes have shown no effect on cognitive function of these products after a single dose.

Claims (19)

The use of a composition comprising a mixture of molecules obtained from Vitis vinifera and Vaccinum angustifolia, wherein the mixture is:
-With at least 1% catechin and / or epicatechin, the percentage is given by weight to the total weight of the mixture.
-With at least 5 ppm (1 million percent in the mixture) ferulic acid,
-At least 200 ppm resveratrol,
Included as a single dose taken on a non-repetitive basis to improve or maintain cognitive function in healthy humans or healthy animals, said single dose taken less than 5 hours prior to the desired effect. Use. The use according to claim 1, wherein the single dose is taken less than 3 hours before the desired effect. Use according to claim 1 or 2 to improve or maintain memory and / or executive function and / or attention and / or concentration and / or learning and / or flexibility and / or planning and / or agility. .. The use according to any one of claims 1 to 3 for improving or maintaining the performance of the healthy human in a test or test requiring sustained cognitive ability in a healthy human. 1. The mixture of molecules used is a mixture of molecules containing at least 5% catechin and / or epicatechin, wherein the percentage is given by weight relative to the total weight of the mixture. Use as described in item 1 of 4. The invention according to any one of claims 1 to 5, wherein the mixture of molecules used is a mixture of molecules containing at least 10 ppm (parts per million in the mixture) of ferulic acid. use. The mixture of molecules used is
-Extracts of Vitis vinifera and / or extracts of Vaccinium angustifolia, and
-The use according to one of claims 1-6, characterized in that it is a mixture of molecules consisting of extracts obtained from Vitis vinifera and Vaccinium angustifolia. The extract of the Vitis vinifera present in the mixture of molecules used is characterized by having a flavanol polymer content of less than 0.5% based on the total weight of the polyphenols in the extract. , The use according to one of claims 1 to 7. The mixture of molecules used is
1. A mixture of molecules comprising malvidin 3 glucoside at a concentration of at least 300 ppm, and / or-quercetin and / or quercetin glycoside at least 50 ppm, and / or-anthocyanidin at least 500 ppm. Use as described in item 1 of 8. The composition comprising said mixture of molecules gives humans or animals-at least 100 μg catechin and / or epicatechin per kg body weight, and / or-at least 0.05 μg ferulic acid per kg body weight, and / or-1 kg body weight. The use according to any one of claims 1-9, characterized in that it is taken in a single dose capable of providing at least 1 μg of resveratrol per. The composition comprising said mixture of molecules gives humans or animals-at least 500 μg catechin and / or epicatechin per kg body weight, and / or-at least 0.25 μg ferulic acid per kg body weight, and / or-1 kg body weight. The use according to any one of claims 1-10, characterized in that it is ingested in a single dose capable of providing at least 5 μg of resveratrol per. The composition comprising a mixture of the molecules in humans or animals-at least 1 mg catechin and / or epicatechin per kg body weight, and / or-at least 0.5 μg ferulic acid per kg body weight, and / or-1 kg body weight. The use according to any one of claims 1 to 11, characterized in that it is taken in a single dose capable of providing at least 10 μg of resveratrol per. The composition comprising said mixture of molecules is ingested in a single dose which allows humans or animals to provide-at least 0.2 μg quercetin per kg body weight and / or-at least 1 μg anthocyanidin per kg body weight. The use according to claim 9, characterized in that. The composition comprising a mixture of the molecules is ingested in a single dose that allows humans or animals to provide-at least 1 μg quercetin per kg body weight and / or-at least 5 μg anthocyanidins per kg body weight. The use according to claim 9, characterized in that. The composition comprising a mixture of the molecules is taken in a single dose that allows humans or animals to provide-at least 0.1 μg quercetin per kg body weight and / or-at least 10 μg anthocyanidins per kg body weight. The use according to claim 9, characterized in that. The composition comprising said mixture of molecules is in a form selected from tablets, capsules, gelatin capsules, powders, solutions, microcapsules, suspensions, emulsions, food supplements, beverages and foods for humans or animals. The use according to any one of claims 1 to 15, characterized in that. The use according to any one of claims 1 to 16 in healthy humans over 10 years of age. The use according to any one of claims 1-16 in healthy animals selected from dogs, cats, and horses. A composition comprising a mixture of molecules obtained from Vitis vinifera and Vaccinum angustifolia, wherein the mixture is:
-With at least 1% catechin and / or epicatechin, the percentage given by weight to the total weight of the mixture.
-With at least 5 ppm (1 million percent in the mixture) ferulic acid,
-At least 200 ppm resveratrol,
Included for a single dose taken on a non-repeating basis to improve or maintain cognitive function in a sick human or sick animal, said single dose less than 5 hours before the desired effect. The composition to be taken.

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